A New 4-Hydroxyphenylcyanide Nitrilases from Pseudomonas sp. and Its Catalytic Properties *

Nitrilases can hydrolyze nitrile efficiently under mild conditions. The enzymatic methods have the advan- tages of less pollution, low cost compared with the chemical methods. Nitrilases are potentially applied in agriculture, industry, environment, and biomedicine. In this study, Berthelot method and high performance liquid chromatography were used to screen new stains for nitrilases. A nitrilase with high substrate specificity for 4-hydroxyphenylcyanide was found from Pseudomonas sp. 6-1. The stain produced 28.47 U/mL nitrilase after optimization of the culture conditions. The enzyme remained 80% activities at pH 6.6 to pH 7.6, temperature 35˚C to 45˚C, and it was stable after 18 h incuba- tion. The stain can be potentially used in biosynthesis 4-Hydroxyphenylacetic acid using 4-hydroxyphenylcyanide as substrate.

[1]  D. Kavan,et al.  Retraction: Heterologous expression, purification and characterization of nitrilase from Aspergillus niger K10 , 2013, BMC Biotechnology.

[2]  V. Křen,et al.  Purification and characterization of heterologously expressed nitrilases from filamentous fungi , 2011, Applied Microbiology and Biotechnology.

[3]  A. Stolz,et al.  Conversion of Sterically Demanding α,α-Disubstituted Phenylacetonitriles by the Arylacetonitrilase from Pseudomonas fluorescens EBC191 , 2011, Applied and Environmental Microbiology.

[4]  D. Wei,et al.  A New Nitrilase-Producing Strain Named Rhodobacter sphaeroides LHS-305: Biocatalytic Characterization and Substrate Specificity , 2011, Applied biochemistry and biotechnology.

[5]  Ludmila Martínková,et al.  Genome mining for the discovery of new nitrilases in filamentous fungi , 2011, Biotechnology Letters.

[6]  Li Zhou,et al.  A high-throughput screening strategy for nitrile-hydrolyzing enzymes based on ferric hydroxamate spectrophotometry , 2011, Applied Microbiology and Biotechnology.

[7]  R. Chaloupkova,et al.  Purification and characterization of nitrilase from Fusarium solani IMI196840 , 2010 .

[8]  Ludmila Martínková,et al.  Biotransformations with nitrilases. , 2010, Current opinion in chemical biology.

[9]  Jung-Kul Lee,et al.  A positively charged amino acid at position 129 in nitrilase from Rhodococcus rhodochrous ATCC 33278 is an essential residue for the activity with meta‐substituted benzonitriles , 2010, FEBS letters.

[10]  S. Kitson,et al.  Hydrolysis of ( 14 C)-Nitrile using Nitrilase (NIT) Biocatalysts , 2010 .

[11]  Huimin Yu,et al.  Gene Cloning, Overexpression, and Characterization of the Nitrilase from Rhodococcus rhodochrous tg1-A6 in E. coli , 2010, Applied biochemistry and biotechnology.

[12]  S. Shimizu,et al.  Versatile nitrilases: Nitrile‐hydrolysing enzymes , 1994 .

[13]  H. Ohta,et al.  Microbial Hydrolysis as a Potent Method for the Preparation of Optically Active Nitriles, Amides, and Carboxylic Acids. , 1992 .